Trng i hc K Thut Cng Ngh TP.HCM Khoa Cng ngh Thc Phm B MN Cng Ngh Thc Phm

N MN HCCng Ngh Thc Phm (MSMH: 20471014)

TI: ANTHOCYANIN V NHNG NGUYN LIU CHA ANTHOCYANIN

GVHD: C Trn Th Cc Phng SVTH: 1. Hunh Th Thanh Tuyn_0851100344 2. Trn Th Xun Hng_ 0851100091 Ngnh: Cng Ngh Thc Phm

Nm hc: 2011

Mc lcI. Tng quan v Anthocyanin...............................................................................................3 1. Gii thiu........................................................................................................................ 4 2. Cu trc ha hc ca Anthocyanin................................................................................. 4 3. Tnh cht ca Anthocyanin .............................................................................................5 Cng dng .......................................................................................................................7 4. S phn b ca Anthocyanin.......................................................................................... 8 5. Qa trnh sinh tng hp Anthocyanin .............................................................................8 5.1. Con ng sinh tng hp Anthocyanin trong t bo thc vt .....................................9 5.2. Cc yu t nh hng n qu trnh sinh tng hp .....................................................9 6. S chuyn i cu trc ca Anthocyanin trong mi trng lng ..................................9 7. Mt s yu t nh hng n mu ca Anthocyanin..............................................11 7.1. Cu trc ......................................................................................................................12 7.1.1 Cu trc chuyn ha .................................................................................................13 7.1.2. Cu trc ha hc .....................................................................................................14 7.2. PH...............................................................................................................................15 7.3. Nhit ......................................................................................................................16 7.4. Oxy .............................................................................................................................17 7.5. Enzyme ......................................................................................................................19 7.6. nh sng ....................................................................................................................20 7.7. ng v cc sn phm bin tnh ca chng .............................................................22 7.8. Cc ion kim loi .........................................................................................................25 7.9. Sunphurdioxide(SO2) ................................................................................................ 28 7.10. Acid Ascorbic ..........................................................................................................30

8. Kh nng ng dng v cc ngun nguyn liu trong sn xut Anthocyanin hin nay.. 34 9. Vai tr ca cc hp cht Anthocyanin ..........................................................................37 9.1. i vi thc vt .........................................................................................................40 9.2. i vi sc khe con ngi .......................................................................................12 9.2.1. Hot tnh chng oxy ha .........................................................................................13 9.2.2. Hot tnh chng ung th .........................................................................................14 9.2.3. Hot tnh chng cc bnh tim mch.........................................................................15 Kt lun ..................................................................................................................16 II. Cc nguyn liu c cha Anthocyanin .........................................................................17 1. Bp ci tm ....................................................................................................................19 1.1. Tng quan ..................................................................................................................20 1.2. Thnh phn ha hc....................................................................................................22 Gii thiu v phng php trch ly .............................................................................25 Cc yu t nh hng n qu trnh trch ly .............................................................. 28 1.3. Quy trnh cng ngh chit tch Anthocyanin t bp ci tm .....................................30 Tin hnh th nghim ..................................................................................................14 Phng php th nghim..............................................................................................15 Mc ch th nghim....................................................................................................16 Chun b th nghim ....................................................................................................17 B tr th nghim .........................................................................................................19 Phng php nghin cu ............................................................................................20 Kt qu nghin cu......................................................................................................22 1.4. Kt lun ......................................................................................................................25 2. Go np than .................................................................................................................19 2.1. Tng quan ..................................................................................................................20 2.2. Thnh phn ha hc....................................................................................................22

2.3. Qu trnh v tinh ch Anthocyanin t go np than ..................................................30 2.4. Nhn xt v kt lun...................................................................................................15 3. Du ty ..........................................................................................................................19 3.1. Tng quan ..................................................................................................................20 3.2. Thnh phn ha hc....................................................................................................22 3.3. Phng php nghin cu ...........................................................................................30 3.3.1. Nguyn liu .............................................................................................................14 3.3.2. Phng php...........................................................................................................15 3.3.3. Tin hnh th nghim...............................................................................................16 3.3.4. Kt qu nghin cu v cc c tnh ca Anthocyanin ............................................17 Nhn xt....................................................................................................................16 3.4. Kt lun ......................................................................................................................17 III. Cc cht Anthocyanin thng mi..............................................................................17 IV. Phng php nghin cu, bo v mu ca Anthocyanin .......................................19 1.Phng php nghin cu................................................................................................20 S nghin cu............................................................................................................22 2. Bo v mu ca Anthocyanin...................................................................................22 Mc Lc_Bng: Bng 1.Cc Anthocyanin trong mt s ngun thc vt.....................................................17 Bng 2. Thnh phn ha hc ca bp ci ..........................................................................19 Bng 3. nh hng ca dung mi n hm lng v mu ca Anthocyanin..............20 Bng 4. T l dung mi v hm lng Anthocyanin.........................................................17 Bng 5. Thnh phn ha hc ca go np than .................................................................19 Bng 6. Thnh phn dinh dng ca qu du ty..............................................................20 Bng 7. Kt qu hm lng Anthocyanin theo chn ca qu du.................................17 Bng 8. Phn trm mu cn li theo thi gian 95oC ti cc PH khc nhau ca

Anthocyanin th chit trong dung mi nc Sulfite v etanol/H2O ................................19 Mc Lc_Hnh: Hnh 1. Cu trc ca mt s Anthocyanin t nhin...........................................................17 Hnh 2. Con ng sinh tng hp Anthocyanin ...............................................................19 Hnh 3. S chuyn ha cu trc ca phn t Anthocyanin trong mi trng lng...........20 Hnh 4. Cu trc chuyn ha ca Anthocyanin trong nc...............................................17 Hnh 5. nh hng ca PH ln tc thy phn ca qu du c un nng ti 45oC trong trng hp c Oxygen hoc Nitrogen ..............................................................................19 Hnh 6. S bin tnh ca Anthocyanin 3,5_diglucoside ti PH 3,7...................................20 Hnh 7. S bin tnh ca Anthocyanin vi phn ng oxy ha Catechol............................20 Hnh 8. Phn ng ngng t ca a) Cyanidin v furfural v b) cyanidin ketobase v furfural...............................................................................................................................17 Hnh 9. S Jurd i vi phn ng thun nghch gia SO2 va Anthocyanin ................19 Hnh 10. S chuyn ha Malvin thnh Malvone bi H2O2 to thnh s oxy ha vitamin C............................................................................................................................20 Hnh 11. Fla-2-ene c to thnh t phn ng gia vitamin C v A...............................17 Hnh 12. S to phc gia Cyanidin v DNA ..................................................................19 Hnh 13. Bp ci tm..........................................................................................................20 Hnh 14. Cng thc cu to ca Cyanidin 3,5_diglucoside...............................................20 Hnh 15. Qa trnh trch ly mt on.................................................................................20 Hnh 16. Qa trnh trch ba on giao dng.......................................................................17 Hnh 17. Qa trnh trch nhiu on nghch dng ............................................................19 Hnh 18. S quy trnh cng ngh chit tch Anthocyanin t bp ci tm....................20 Hnh 19. th biu din nh hng ca cc h dung mi...............................................20 Hnh 20. nh hng ca t l dung mi (Etanol-nc ) n hm lng Anthocyanin . . .19 Hnh 21. Ht np than........................................................................................................20

Hnh 22. Qa du ty.........................................................................................................20 Hnh 23. Kt qu hm lng Anthocyanin cc nng SO2 khc nhau .......................20 Hnh 24. Kt qu hm lng Anthocyanin cc nhit khc nhau ..............................20

I. Tng quan v Anthocyanin:1. Gii thiu: Cc Anthocyanin hin thuc nhm cc cht mu t nhin tan trong nc ln nht trong th gii thc vt. Thut ng anthocyanin bt ngun t ting Hy Lp, trong Anthocyanin l s kt hp gia Anthos ngha l hoa v Kysanesos ngha l mu xanh. Tuy nhin, khng ch c mu xanh, anthocyanin cn mang n cho thc vt nhiu mu sc rc r khc nh hng, , cam v cc gam mu trung gian. Cc anthocyanin khi mt ht nhm ng c gi l anthocyanidin hay aglycon. Mi anthocyanidin c th b glycosyl ha acylate bi cc loi ng v cc acid khc ti cc v tr khc nhau. V th lng anthocyanin ln hn anthocyanidin t 15-20 ln. 2. Cu trc ha hc ca Anthocyanin: Anthocyanin thuc nhm cc hp cht flavonoid, c kh nng ha tan trong nc v cha trong cc khng bo. V bn cht, cc Anthocyanin l nhng hp cht glycoside ca cc dn xut polyhydroxy v polymethoxy ca 2-phenylbenzopyrylium hoc mui flavylium. Cho n nay, ngi ta xc nh c 18 loi aglycon khc nhau, trong 6 loi ph bin nht l pelargonidin, cyanidin, delphinidin, peonidin, petunidin v maldivin. Trong t nhin, Anthocyanin rt him khi trng thi t do (khng b glycosyl ha). Nhm hydroxy t do v tr C-3 lm cho phn t anthocyanidin tr nn khng n nh v lm gim kh nng ha tan ca n so vi anthocyanin tng ng. V vy, s glycosyl ha lun din ra, u tin v tr nhm 3-hydroxy. Nu c thm mt phn t ng sa, v tr tip theo b glycosyl ha thng gp nht l C-5. Ngoi ra, s glycosyl ha cn c th gp cc v tr C7, C-3, C-5. Loi ng ph bin nht l glucose, ngoi ra cng c mt vi loi monosaccharide (nh galactose, rammose, arabinose), cc loi disaccharide (ch yu l rutinose, sambubiose hay sophorose) hoc trisaccharide tham gia vo qu trnh glycosyl ha. S methoxyl ha cc anthocyanin v cc glucoside tng ng din ra thng thng nht l v tr C-3 v C-5, cng c th gp v tr C-7 v C-5. Tuy nhin, cho n nay, ngi ta vn cha tm thy mt hp cht no b glycosyl ha hay b methoxyl ha trn tt c cc v tr C-3,

-5, -7 v -4 do cn thit phi cn t nht mt nhm hydroxyl t do C-5, -7 hay -4 hnh thnh dng cu trc quinonoidal base (dng cu trc ca anthocyanin thng tn ti trong khng bo thc vt c pH t 2,5 7,5).

Hnh 1: Cu trc ca mt s Anthocyanidin t nhin. Cc Anthocyanin tng ng lunc glycosyl ha nhm hydroxy C-3 S acyl ha cng c th xy ra v tr C-3 ca phn t ng hay ester ha nhm hydroxy C-6. Cc nhm acyl ha chnh l cc phenolic acid nh -coumeric, caffeic, ferulic hay sinapic acid v mt lot cc acid nh acetic, malic, malonic, axalic v succinic. 3. Tnh cht ca Anthocyanin: Anthocyanin tinh khit dng tinh th hoc v nh hnh l hp cht kh phn cc nn tan tt trong dung mi phn cc. Anthocyanin ha tan tt trong nc v trong dch bo ha. Khi kt hp vi ng l cho phn t anthocyanin ha tan hn. Mu sc anthocyanin thay i ph thuc vo nhit , cc cht mu v nhiu yu t khc, Khi tng s lng nhm OH trong vng benzene th mu cng xanh m. Mc methyl ha cc nhm OH vng benzene cng cao th mu cng . Nu nhm OH v tr th ba kt hp vi cc gc ng th mu sc cng s thay i theo s lng cc gc ng c nh vo nhiu hay t.

Cc Anthocyanin cng ph thuc rt mnh vo pH ca mi trng: Khi pH > 7 cc Anthocyanin c mu xanh v khi pH < 7 cc Anthocyanin c mu . pH = 1 cc Anthocyanin thng dng mui oxonium mu cam n . pH = 45 chng c th chuyn v dng baz Cacbinol hay baz Chalcon

khng mu. pH = 78 li v dng baz Quinoidal Anhydro mu xanh.

Mu sc ca anthocyanin cn c th thay i do hp th trn polysaccharide. Khi un nng lu di cc anthocyanin c th ph hy v mt mu. Anthocyanin c bc sng hp th trong min nhn thy, kh nng hp th cc i ti bc sng 510540nm. hp th l yu t lin quan mt thit n mu sc ca cc anthocyanin chng ph thuc vo pH ca dung dch, nng anthocyanin: thng pH thuc vng acid mnh c hp th ln, nng anthocyanin cng ln hp th cng mnh. Tm li, trong mi trng acid, cc anthocyanin l nhng baz mnh v c th to mui bn vng vi acid. Anthocyanin cng c kh nng cho mui vi baz. Nh vy chng c tnh cht amphote. Mui vi acid th c mu , cn mui vi kim th c mu xanh. Cng dng: Anthocyanin l cht mu thin nhin c s dng kh ph bin v an ton trong thc phm, to ra nhiu mu sc hp dn cho mi sn phm. Anthocyanin l hp cht c nhiu hot tnh sinh hc qu nh:

Kh nng chng oxy ha cao nn c s dng chng lo ha, hoc chng oxy ha cc sn phm thc phm, chng vim, chng cc tia phng x Hn ch s suy gim sc khng, s pht trin ca cc t bo ung th. Ngoi ra, Athocyanin cn c vai tr thu ht cn trng gip cho s th phn ca cy din ra tt hn.

4. S phn b ca Anthocyanin: anthocyanin tp trung nhng cy ht kn v nhng loi ra hoa, phn ln nm hoa v qu, ngoi ra cng c l v r. Trong nhng loi thc vt ny, anthocyanin c tm thy ch yu cc lp t bo nm bn ngoi nh biu b.

Cc hp cht anthocyanin xut hin rng ri trong khong t nht 27 h, 73 loi v trong v s ging thc vt s dng lm thc vt (Bridle v Timberlake, 1996). Cc h thc vt nh vitaceae (nho) v rosaceae (cherry, du ty, mm xi, to,) l cc ngun anthocyanin ch yu. Bn cnh cn c cc h thc vt khc nh solanceae ( c tm), saxifragaceae (qu l v en), ericaceae (qu vit quc) v brassicaceae (bp ci tm). Cc loi anthocyanin ph bin nht l cc glucoside ca cyanidin, k n l pelargonidin, peonidin v delphinidin, sau petuidin v maldivin. S lng cc 3glucoside nhiu gp 2,5 ln cc 3,5glucoside. Loi anthocyanin hay gp nht chnh l Cyaidin-3-glucoside. Bng 1: Cc Anthocyanin trong mt s ngun thc vt Tn thc vt Allium cepa Tn thng thng C hnh Loi Anthocyanin Cy-3-glucoside, 3-galactoside, 3diglucoside v 3-laminarriobioside, Pn-3-glucoside Cy-3-sophoroside-5-glucoside cacyl ha vi malonoyl, -coumaroyl, di-coumarol,feruloyl, diferuloyl, sinapoyl v disinapoyl Pg v Cy-3-glucoside Cy v Dp-3-glucoside Cy, Pn, mono- v biosides Pg v Cy-3-sophoroside-5-glucoside acyl ha vi -coumaroyl, feruloyl, caffeoyl. Cy, Pn, Dp, Pt v Mv mono v diglucoside; dnh t do v dng acyl ha

Brassica oleraea

Bp ci tm (red cabbage)

Fragaria spp Glycine maxima Hibicus sabdariffa L Raphanus sativus

Du ty (strawberry) u nnh (v) Hoa bt dm C ci (r)

Vitis spp

Nho

Cy = cyanidin, Pg= pelagorindin, Pn= peonidin, Dp= delphinidin, Pt = petunidin, Mv= malvidin

5.

Qu trnh sinh tng hp Anthocyanin:

5.1. Con ng sinh tng hp Anthocyanin trong t bo thc vt: Anthocyanin thng nm trong cc m hoa v qu, trong cc lp t bo biu b v di biu b ca l v thn thc vt. Trc y, c ngi cho rng qu trnh sinh tng hp anthocyanin din ra trong cc bo quan hnh cu nm bn trong khng bo, c gi l cc anthocyanoplast. Cc bo quan ny xut hin rng ri c cc cy mt l mm v cy hai l mm. Tuy nhin, theo nghin cu cho thy qu trnh sinh tng hp anthocyanin c th khng din ra trong cc anthocyanoplast do y c hm lng anthocyanin cao v pH thp, trong khi pH ti thch ca tt c cc enzyme c bit tham gia vo con ng sinh tng hp flavonoid l pH trung tnh. Qu trnh sinh tng hp anthocyanin c iu ha bi mt h thng cc gen. Anthocyanin ch xut hin cc loi thc vt trn cn m khng c cc loi ng vt, vi sinh vt hay thc vt di nc. Nguyn nhn do qu trnh sinh tng hp anthocyanin i hi phi c vt liu khi u i t qu trnh quang hp. Di tc dng ca emzyme Chalcone synthase, Chalcone c to thnh thng qua phn ng ngng t gia 1 phn t acid cinammic hot ha (coenzyme A ca 4-coumaric acid hay t gp hn l ca ferulic, caffeic, 5-hydroxy ferulic hay sinapic acid) v 3 phn t malonyl-CoA. Acid cinammic hot ha c ly t L-phenylalanine qua con ng sinh tng hp phenylpropanoid(gm cc enzyme Phenylalanine-ammonialyase, Cinammate-4-Hydroxylase, 4-Coumarat-CoA ligase). Malonyl-CoA c to thnh t AcetylCoA qua phn ng xc tc bi enzyme Acetyl-CoA carboxylase. Tip theo, phn t Chalcone c ng vng thng qua phn ng xc tc bi enzyme Chalcone isomerase hnh thnh cc flavanone ng phn (nh naringenin). Sau , chnh cc hp cht ny c chuyn ha thnh cc dihydroxyflavanol (flanva-3-ol) bi enzyme flavanone-3-hydroxylase, ng thi c th kt hp vi c enzyme 2-hydroxylase v mt enzyme dehydratase. Cc dihydroxyflavonol c xem l cc tin cht ca anthocyanidin. Cc enzyme xc tc cho qu trnh chuyn ha t cc dihydroxyflavonol thnh cc anthocyanidin cho n nay vn cha c xc nh r rng, c th l cc oxydo-reductase vi cc hp cht trung gian l flavan-3,4-diol (leucoanthocyanidin). S khc nhau nhm th vng B c th bt u giai on cinammic acid, nhng ch yu din ra giai on hnh thnh cc hp cht trung gian C15 (nh chalcone/flavanone) sau . Cc hp cht anthocyanidin t nhin s c glycosyl ha ngay lp tc nhm hydroxy C-3 do chng khng bn trong mi trng lng. Qu trnh glycosyl ha tip theo, nu c, s xy ra theo dng bc thang, bt u t vic gn cc phn t ng vo gc 3-glycosyl v hoc nhm 5-hydroxy. Cc uridine diphosphat (UDP)-ng ng vai tr l cht cho glycosyl cho tt c cc

enzyme glycosyl-transferase. Qu trnh acyl ha Anthocyanin c th xy ra sau qu trnh glycosyl ha, xc tc bi cc enzyme acyltransferase. 5.2. Cc yu t nh hng n qu trnh sinh tng hp: Qu trnh sinh tng hp anthocyanin b tc ng bi cc yu t mi trng nh: nh sng, nhit , cc hormone thc vt, ngun dinh dng, nhng tc ng c hc v s tn cng ca cc loi gy bnh. Trong , nh sng chnh l yu t quan trng nht. nh sng hng ngoi hot ha cc phytocrome mu, l cht cm ng cho cc enzyme trong con ng flavone-glucoside v t , gip tng hp anthocyanin trong t bo. Ngoi ra, cng thng cn n tia UV bn cnh cc phytocrome hot ng cm ng cc enzyme. Trong mi trng nui cy t bo Haplopappus gracilis, ch c tia UV c bc sng nh hn 345nm mi kch thch c qu trnh tng hp anthocyanin. Di tc dng ca tia UV, hot tnh ca cc enzyme phenylalanineammonia-lyase, chalcone, synthase v chalcone isomerase tng ln ng k. Qu trnh tch t Anthocyanin trong l cy cn lin quan n mc pht trin ca l, s thiu ht cc cht khong a lng nh N, P, mc tip xc ca l vi tia UV. Ngoi ra, s tng hp anthocyanin cng c th l mt phn ng ca thc vt chng li s tn cng ca cc loi n c cng nh ca cc loi nm mc.

Hnh 2: Con ng sinh tng hp Anthocyanin. Enzyme: chalcone synthase (a); chalcone isomerase (b); flavanone 3 hydroxylase (c); oxidoreductase (s) (d); glycosyl transferase (e) Gl = nhm glycosyl; R, R=H, OH, OCH3 hay OGl6. S chuyn i cu trc ca Anthocyanin trong mi trng lng:

Trong mi trng lng, di tc ng ca cc gi tr pH khc nhau ca mi trng, cu trc ca phn t Anthocyanin s tri qua mt s chuyn ha. mt gi tr pH xc nh, din ra s cn bng gia 4 cu trc Anthocyanin: Quinonoidal (anhydro) base (A): mu hi tm Cation flavylium (AH+): mu Carbinol pseudobase (hay hemiacetal) (B): khng mu Chalcone : khng mu

Phng trnh chuyn ha: AH+ A + H+

AH+ + H2O

B + H+

B

C

S chuyn i cu trc ca Anthocyanin trong nc l do hot tnh cao ca gc aglycone (Anthocyanidin). Gc ng, nhm methoxyl cng nh cc nhm acyl ha cng nh hng ng k n s chuyn ha trn.

Hnh 3: S chuyn ha cu trc ca phn t Anthocyanin trong mi trng lng Dng quinonoidal base c to thnh thng thng pH > 3. Cation flavylium tng i bn mi trng acid (nh pH < 3). Trong mi trng trung tnh hay acid yu, Anthocyanin tn ti ch yu cc dng khng mu. V vy, s n nh ca cc dng mang mu, c bit l dng quinonoidal base, mt phn l do s xut hin ca cc gc acid gn vo phn t ng, hay cn gi l s co-pigment ha ni phn t. S d s co-pigment ha ni phn t gip tng bn ca phn t anthocyanin l do s xp chng ca vng thm trong gc acid vi vng pyrylium ca phn t anthocyanin, gim bt kh nng b cc phn t nc tn cng hnh thnh cc hp cht carbinol, chalcone khng mu. Ngoi ra, hiu ng copiment ha ngoi phn t cng c th gip lm bn cc cu trc quinoidal base hay ion flavylium do s c mt ca nhng phc cht khng mu, nh cc flavone, flavonol. Trong qu trnh x l nhit v nh sng, qu trnh copigment ha

ngoi phn t gi vai tr quan trng, ngn chn s mt mu c th xy ra. C ch phn hy do nhit ni chung do s chuyn ha thun nghch ca cc ion flavylium thnh cc dng chalcone. Chalcone l cht trung gian trong c hai qu trnh phn hy do nhit v nh sng ca anthocyanin. Theo cc nghin cu gn y (Davis v cng s, 1983; Mazza v Brouillard,1990) hiu ng co-pigment ha chnh l c ch lm bn mu chnh ca anthocyanin trong thc vt do n lm gim t l gia dng khng mang mu (chalcone) v dng mang mu thng qua vic to phc vi dng mang mu, ngn chn qu trnh chuyn ha t dng mang mu sang dng khng mang mu. Trong dung dch, Anthocyanin dng khng acyl ha hay acyl ha mt n v ng vai tr nh mt cht ch th pH, tn ti dng acid hay base ty thuc vo gi tr pH. pH acid (pH < 3), dung dch anthocyanin cho mu m. Khi pH tng, mu sc ca dung dch s nht dn v chuyn sang khng mu v cui cng c mu tm hay xanh dng pH cao (pH > 6).6. Mt s yu t nh hng n mu ca Anthocyanin:

So vi a s cc cht mu thin nhin, anthocyanin l cht mu c bn km hn, n ch th hin tnh bn trong mi trng acid. Ngoi ra, n c th phn hy to thnh dng khng mu v sn phm cui cng ca s phn hy c dng mu nu cng vi nhng sn phm khng tan.

S phn hy anthocyanin c th xy ra trong qu trnh trch v tinh ch chng, ng thi s phn hy ny cn xy ra trong qu trnh x l v bo qun cc sn phm thc phm. bn ca cc anthocyanin ph thuc vo nhiu yu t nh: cu trc ha hc ca anthocyanin, pH, nhit , s c mt ca copigment, ion kim loi, oxy, acid ascorbic, SO2, nh sng, enzyme, ng v cc sn phm bin tnh ca chng.

7.1. Cu trc: 7.1.1.Cu trc chuyn ha: Trong mi trng nc cc anthocyanin t nhin ging nh cht ch th pH. pH thp, xanh pH trung gian, v khng mu pH cao. Ti pH cho, tn ti mt cn bng gia 4 cu trc ca anthocyanin v aglycone: quinonoidal (anhydro) bazo (A) mu xanh, cation flavylium (AH+ ) mu , carbinol pseudobase (B), v chalcone (C) khng mu.

Hnh 4: Cu trc chuyn ho ca anthocyanin trong nc Khi pH < 2.0, cc anthocyanin tn ti ch yu dng cation flavylium mu (R3=0-ng) hoc mu vng (R3=H). Khi pH tng, s mt prton xy ra nhanh thnh dng quinononidal mu xanh dng hoc mu . Dng quinonoidal thng tn ti nh mt hn hp v pKa ca nhm OH v tr 4,7 v 5 l tng t. Khi yn, s chuyn ha xy ra : c s tch nc ca cation flavylium cho ra dng pseudobase (hemiacetal), Dng ny cn bng vi dng chalcone vng m khng mu. Hai dng Cis (CE) v transchalcone(CZ) c to thnh t carbinol pseudobase bi phn ng m vng nhanh v isomer ha chm. C 2 chalcone ny khc nhau so vi chalcone bnh thng l bi chng c nhm chc carbonyl cnh vng B, trong chalcone bnh thng c nhm carbonyl k cn vng A. i. Cu trc ha hc: bn mu v cng mu ca cc anthocyanin ph thuc vo v tr v s lng ca cc nhm hydroxyl, methoxyl, ng v cc ng c acyl ha. Khi s nhm hydroxyl trong vng B tng,cc i hp thu vng thy c dch chuyn v pha c bc sng di hn v mu thay i t cam n xanh dng. V d: Bc sng hp thu cc i trong dung dch HCL 0,01% MeOH i vi: pelgonidin: max

= 520 nm (cam)

cyaniding: = 535 nm ( cam) max dephinidin: max

= 545 nm ( xanh)

Khi nhm methoxyl thay th nhm hydroxyl th ta thu c kt qu ngc li. Nhm hydroxyl ti v tr C-3 c ngha quan trng v dung dch Anthocyanin chuyn t mu cam vng n mu , iu ny gii thch s khc nhau gia Anthocyanin c mu trong khi 3 deoxyanthocyanin : apigenidin, luteolinidin v tricetinidin c mu vng nhng 3 deoxyanthocyanidin bn hn cc anthocyanidin khc (Mazza v Brouillard, 1987; Iacobucci v Sny, 1983). S c mt ca nhm hydroxyl ti v tr C-5 v nhm th v tr C-4, c 2 bin ha dng c mu thng qua s ngn cn cc phn ng hydrat ha n n s to thnh dng khng mu. Khi mc hydroxyl ha cc aglycone tng, tnh bn ca cc anthocyanin s gim. Tuy nhin khi tng s methoxyl ha, s thu c kt qu ngc li. V d: s c mt ca nhm OH v tr 4 v 7 trong phn t lm bn ha ng k cc pigment, trong khi , s methoxyl ha c nhm hydroxyl ny lm gim bn. Cc anthocyanin c glycosyl ha v acyl ha s cho dng mu xanh, S glycol ha nhng nhm OH t do lm tng tnh bn ca anthocyanin. V vy, cc diglucoside bn hn cc monogluside ca cng mt anthocyanin. Anthocnin c cha 2 hay nhiu nhm acyl (nh ternatin, platyconin, cinerarrin, gntiodenphin v zebrrinin) l bn trong mi trng trung tnh hoc acid yu do lin kt hydro gia cc nhm hydroxyl ca cc nhn phenolic trong anthocyanin v acid vng thm. Brouillard (1981-1982) v Goto cng vi cng vin (1982-1983) kho st rng cc anthocnin diacylate ha c bn ha bi s lin kt cht kiu sandwich nh s tng tc gia vng anthocyanin v 2 nhm acyl vng thm. 7.2. PH: Trong mi trng nc, pH c nh hng ng k ln mu sc ca anthocyanin (Brouillard 1984,Mzza v Brouillard 1987). Cu trc, bn mu, mu sc ca anthocyanin thay i theo s thay i ca ph. S thay i cu trc ca anthocyanin khi pH thay i c cp trong phn cu trc chuyn ha.

Nm 1956 Lukton nghin cu thy rng: ph = 2 4.5 th vn tc phn hy pelargonidin 3- glucoside trong dch p ca qu du khng xc nh c khi khng c O2. Khi c mt O2 ph tng nhanh, s bin tnh tng ln mnh lit.

Hnh 5: nh hng ca pH ln tc thy phn ca qu du c un nng ti 450C trong trng hp c Oxygen hoc Nitrogen

n nm 1972, Adams nghin cu thy rng ph = 2 4 t nh hng n s phn hy ca Anthocyanin trong sut qu trnh gia nhit khi khng c O2. Khi c mt O2 th Anthocyanin b bin tnh mnh ph trn cu trc ca chng cng vi bn mu thay i vi s thay i ph. 7.3. Nhit :

Trong qu trnh ch bin v bo qun thc phm, Anthocyanin d dng b bin tnh di tc dng ca nhit (Markakis 1974).

Hnh 6: S bin tnh ca Anthocyanin 3,5 di Glucoside ti pH 3,7

Khi mt cu trc ca anthocyanin bn vi s gia tng ca pH th n cng bn vi s gia tng ca nhit . S hydroxyl ha cc aglycone lm gim tnh bn ca anthocyanin, trong khi s methxyl ha, glycosyl ha, acyl ha s cho kt qu ngc li. V d: Anthocyanin 3- glycoside c bn nhit ln nht ti pH = 1.8 - 2.0 vi s c mt ca oxy trong khi anthocyanidin 3,5-diglycoside c bn nhit ln nht ti pH = 4.0 5.0 C ch ca s phn hy nhit xy ra khng nhng ph thuc vo nhit m cn ph thuc vo bn thn anthocyanin . Drazdina cho rng coumarin glycoside l sn phm phn hy ph bin ca anthocyanin 3,5 diglycoside v c ch ca s phn hy l: u tin, cation flavylium chuyn thnh dng quinnidal base, tip theo l hnh thnh nhiu dng sn phm trung gian v cui cng thu c dn xut coumarin v thnh phn tng ng vi vng B ca anthocyanidin, S bin tnh ny khng nhng c xc tin bi nhit m cn b nh hng bi oxy. Anthocyanidin 3 glycoside th khng hnh thnh nhng dn xut coumrin m bc u tin ca s phn hy bao gm s chuyn ha ca dng carbinol pseudobase khng mu, sau l s m vng pyrylium hnh thnh dng chalcone trc khi thy phn lin kt glycoside.

Adams cho rng, cc anthocyanidin 3-glycoside khi c nung nng pH = 2.0 4.0 u tin s b thy phn lin kt glycoside ( 100oC) sau l s bin i aglycone thnh chalcone. S bin tnh hn na dn n hnh thnh dng sn phm mu nu c bit l vi s c mt ca oxy. Khi un nng lu di cc anthocyanin c th b phn hu v mt mu, c bit l cc anthocyanin ca dy ty, anh o, c ci. Ngc li cc anthocyanin ca phc bn t en cng trong iu kin li khng b thay i.Nhn chung khi gia nhit, cc cht mu d dng b phn hu, cn cc cht mu vng th kh hn. 7.4. Oxy: Oxy v nhit c xem l nhng tc nhn c trng xc tin s phn hy ca anthocyanin, do m sinh ra nhng dng sn phm khng mu hoc mu nu. Chnh s oxy ha trc tip dng carbinol pseudobase gy ra s kt ta v ng vng trong nc tri cy. Trong qu trnh un nng cc anthocyanin b oxy ha mnh lit. Oxy v nhit l nhng tc nhn xc tin c bit nht trong nc p ca blueberry, cherry (anh o), currant, nho, raspberry v du. Lng anthocyanin cn li ca du s ln hn khi ng chai di iu kin chn khng hoc nitrogen (Baravingas v Cain, 1965). bn ca cc pigment ca nho cn c s dng nh l cht mu nc gii kht c tng ln khi ng hp vi nitrogen.

7.5. Enzyme Nhiu enzyme ni sinh trong t bo ca cy c kh nng lm mt mu anthocyanin. Nhng enzyme ny c gi chung l anthocyanase. Da vo c tnh ca cc enzyme m ngi ta phn lm 2 nhm; Glycosidase v polyphenol oxidase (PPO). Cc enzyme ny thu c t nm (fugal).

Glycosidase: l enzyme thy phn lin kt glycoside ca anthocyanin to ra ng

t do v aglycone ny km bn hn rt nhiu v mt mu rt nhanh khi c mt ca catecol(Huang 1955).

Polyphenol oxidase (PPO): tc dng ln anthocyanin vi s c mt ca O-diphenol

thng qua c ch oxy ha kt hp. Theo Gromeck v Markakis, s thm vo glycosidase v PPO xc tc cho qu trnh peroxide ha phn hy anthocyanin.

Hnh 7: S bin tnh anthocyanin vi phn ng oxy ho catechol Peng v Markakis ngh mt c ch m trong O-quinine c to thnh bi s oxy ha anthocyanin. Blom phn lp c enzyme anthocyanin-glycosidase t Aspergillus Niger v ch ra nh hng ca n trong qu trnh thy phn lin kt glycoside ca anthocyanin. PPO l enzyme c nh hng yu ln anthocyanin dng quinonoidal base d b thy phn bi PPO hn dng cation flavylium. Tc phn hy bi PPO ph thuc vo s thay th m hnh ca vng B v mc glycosyl ha. 7.6. nh sng Cc anthocyanin thng khng bn khi tip xc vi tia t ngoi, nh sng thy c, v cc ngun php x khc. nh sng c 2 nh hng n anthocyanin: Tng cng cho qu trnh sinh tng hp. Xc tin s bin tnh ca chng.

Nm 1964, Siegenman cho rng nhng qu to ging s chuyn sang mu xanh khi trong bng ti. Nm 1968 Vanburen v cc cng s tng trnh rng cc diglucoside c acyl

ha v methyl ha th cc Anthocyanin trong ru bn nht khi ngoi nh sng, cc diglucoside khng b acyl ha l t bn hn v monoglucoside l km bn nht, nm 1975 Palamidis v Markakis tm thy rng nh sng thc y qu trnh phn hy anthocyanin trong nc gii kht c CO2 c phi mu vi anthocyanin t xc nho. Macccarone v cng s nghin cu s quang ha ca anthocyanin v ch ra rng , anthocyanin diglycosyl ha ti v tr C3 v C-5 l bn hn cc anthocyanin mono glycoyl ha ti v tr C-3 ng thi chng bn hn so vi cc aglycone tng ng. 7.7. ng v cc sn phm bin tnh ca chng: nng 100 ppm, ng v cc sn phm phn hy ca chng c tc dng thc y s phn hy cc anthocyanin, trong fructose, arabinose, lactose v sorbose c kh nng phn hy anthocyanin mnh hn glucose, sucrose, v maltose. Tc phn hy ca anthocyanin lin quan n tc phn hy ca ng. Cc sn phm phn hy ca ng gm c: furfura l,5 hydroxymethyl furfural v acctaldehyde thu c t phn ng Mailard hoc t s oxy ha ca acid ascorbic, polyuronic hoc bn thn cc anthocyanin. Nhng sn phm phn hy ny d dng ngng t vi cc anthocyanin hnh thnh nhng hp cht phc tp c mu nu sm. S phn hy anthocyanin vi s c mt ca furfural v HMF trc tip ph thuc vo nhit v c thy r nht l trong nc tri cy s c mt ca n lm tng thm hiu qu phn hy ca tt c cc loi ung v cc dn xut ca chng.

Hnh 8: Phn ng ngng t ca a) Cyanidin v furfural b) Cyanidin ketobase v furfural

7.8. Cc ion kim loi: Mt s ion kim loi a ha tr c th tng tc vi cc anthocyanin c nhm OH v tr ortho gy ra hiu ng su mu (bathocromic). Hin tng ny xy ra khi kim loi tip xc vi anthocyanin trong qu trnh ch bin rau qu hoc s cho thm cc mui kim loi vo trong thc phm. Sistrunk v Cash chng minh rng c th bn ha mu ca dch trch du bng cch thm vo mui thic. Francis (1977) cng b rng, cc ion Ca, Fe, Al to thm s bo v cho anthocyanin ca nc p tri mn vit qut (cranberry), nhng s bin i mu xy ra l do s to phc gia ion kim loi v tannin, kt qu sau cng l khng c li. Trong cng nghip hp, s mt mu ca nhng tri cy c cha anthocyanin l do c phn ng vi thic ca hp (Culpepper v caldwell, 1972). Trong phn ng vi thic, anthocyanin ng vai tr nh cht kh cc catod hoc anod. Cht kh cc catod c th b kh bi hydro mi sinh t phn ng gia kim loi v acid, cn cht kh cc anod thng l cc anthocyanin c t nht 2 nhm hydroxyl v tr ortho. 7.9. Sunphurdioxide (SO2) Cc anthocyanin thng b mt mu khi c mt ca SO2. Hin tng ny thng xy ra trong qu trnh x l cc sn phm thc phm c cha anthocyanin bng SO2. Qu trnh kh ny c th l thun nghch hoc bt thun nghch. Tri cy v nc qu c x l bng mt lng trung bnh SO2 (500 2000 ppm), lm mt mu cc anthocynin ca chng trc khi ch bin, hn na, chng c desunfit ha v mu anthocyanin c phc hi. Jurd ngh s phn ng thun nghch gia SO2 v anthocyanin trong , dng c mu flavylium phn ng vi ion bisulphate to thnh chromene-2-(hoc 4)- sulphonic acid.

Hnh 9: S Jurd i vi phn ng thun nghch gia SO2 v anthocyanin SO2 nng rt thp (khong 30 ppm) c th c ch s bin tnh do enzyme ca anthocyanin trong qu anh o nhng khng lm mt mu chng (Goodman v Markakis, 1965 ). S ty mu bt thun nghch xy ra trong qu trnh ty qu vi lng ln SO2 (0.8 1.5%) v soda (0.4 1.0%) c dng trong qu trnh ty qu. Phn ng bt thun nghch ny cha c bit hon ton. 7.10. Acid ascorbic Nhiu nh kho st (Beatic v cng s 1943; Pederson v cng s 1947; Kertesz 1952; Meschter 1953; Markakis v cng s 1957; Starr v Francis 1968) quan st s bin mt ng thi ca acid ascorbic v anthocyanin trong nc tri cy tn tr v ngh mt tng tc c th c gia 2 hp cht ny. Acid ascorbic hin din trong hu ht cc sn phm tri cy,vitamin ny b oxy ha to thnh H2O2 v chnh H2O2 lm mt mu anthocyanin.

Hnh 10: S chuyn ho malvin thnh malvone bi H2 O2 to thnh t s oxy ho vitamin C

Shrikhande v Francis tm thy rng cc ion ng xc tin v cc flavonoid lm gim s phn hy ca c hai acid ascorbic v anthocyanin. Nhng sn phm khng mu. Acid dehyroascorbic cng c th lm mt mu anthocyanin nhng ti tc thp hn acid ascorbic.

Hnh 11: Fla-2-ene c to thnh t phn ng gia vitamin C v anthocyanin

8. Kh nng ng dng v cc ngun nguyn liu trong sn xut Anthocyanin hin nay: Cht mu Anthocyanin (E163) c cho php s dng rng ri trong cc loi thc phm nh ung (30 40 ppm), mt tri cy (20 60 ppm), bnh ko (chewing gum, ko cng,

ko mm), yoghurt, cc sn phm trng ming hay cc loi bt ha tan ung lin. Thng thng, hm lng cho php s dng ca Anthocyanin khng b gii hn v ch yu ph thuc vo tng quy trnh sn xut. Hin nay, m bo sc khe ngi tiu dng ngy cng quan tm n vic s dng cc cht mu t nhin hn l s dng cc cht mu tng hp, vn c th gy nhng tc dng xu v sc khe cho ngi s dng. Rt nhiu loi dch trch t cc loi rau v tri c mu sc to bi cc Anthocyanin c s dng to mu cho thc phm. Tuy nhin, cc ngun nguyn liu c la chn sn xut Anthocyanin thng b gii hn bi nng sut thu nhn v gi tr kinh t. Mt trong nhng ngun thu nhn Anthocyanin c hiu qu nht hin nay l cc ngun m trong Anthocyanin ng vai tr l mt ph phm ca qu trnh sn xut cc sn phm c gi tr cao hn; v d nh Anthocyanin t b nho p, b du tm, hay t qu trnh sn xut cht to ngt t nhin Miraculin qu Miracle. Trong , b nho chnh l ngun nguyn liu thu nhn Anthocyanin c hiu qu kinh t v d kim nht. Nho l loi tri cy c sn lng thu hoch hng nm cao nht so vi tt c cc loi tri cy khc. Qu trnh sn xut ru vang v nc p t nho to ra mt lng ln ph phm l b nho. Trong b nho, cc cht mu Anthocyanin chnh l nhng thnh phn c gi tr v c tim nng khai thc nhiu nht. Mu Anthocyanin dng bt sy phun hay dng dch lng c c t nho c a ra th trng t nhiu nm vi tn thng mi l Enocolor v c gii thiu s dng trong thc phm v m phm. Cc ngun thu nhn ph bin khc nh: bp ci tm, go np than, c ci (red radish), khoai lang tm, carrot (black carrot), qu anh o, blackberry v elderberry. Trong , cc ngun nh c ci , khoai lang tm, bp ci tm cung cp hm lng cc Anthocyanin c acyl ha cao hn, do bn mu ca cc dch trch ny cng cao hn. Hin nay, xu hng nghin cu chnh ang tp trung vo vic khai thc cc Anthocyanin c acyl ha c mu sc, bn nhit, ng sng, pH, SO2 tt hn cng vi s pht trin cc k thut nui cy m, t bo c th nng cao kh nng ng dng ca cht mu Anthocyanin trong thc phm. 9. Vai tr ca cc hp cht Anthocyanin: 9.1. i vi thc vt:

Trong gii thc vt, chlorophyll l loi cht mu chim u th v ng mt vai tr rt quan trng trong qu trnh quang hp. S xut hin ca cc cht mu khc, khng phi mu xanh (Anthocyanin, Carotenoid), thng nhm mc ch chnh l to ra s tng phn, thu ht cc loi ng vt, to iu kin cho qu trnh th phn v pht tn ht ging ca cy. Cc nghin cu mi y chng minh c rng cc Anthocyanin gip che ch cc dip lp, bo v cc dip lp chng li cng nh sng cao, gip ngn s c ch qu trnh quang hp. ChalkerScott (1999) ra 3 vai tr chnh ca Anthocyanin trong thc vt gm: Hp th cc tia bc x Vn chuyn cc monosaccharide iu chnh p sut thm thu trong cc giai on kh hn v nhit thp. Ngoi ra, ngi ta cn pht hin Anthocyanin mt s vai tr khc nh: Hale v cng s (2001, 2002) thy rng khi hm lng cht mu tng th kh nng tch t molybden v tungsten loi Brassica sp. (Bp ci) cng tng; hay cc m c cha Anthocyanin, kh nng b tn cng bi nm mc gim i (Coley v Aide,1989). Theo Gould, McKelvie v Markham (2002) Anthocyanin tch t trong t bo l do kt qu ca s tn thng c hc hay do thiu ht phosphoros hoc nitrogen (Chalker-Scott,1999). Ni chung, ngi ta tin rng Anthocyanin c kh nng tng cng phn ng chng oxy ha ca t bo i vi cc yu t gy stress. 9.2. i vi sc khe con ngi: Ngoi nhng vai tr sinh l i vi thc vt, cc hp cht Anthocyanin cn c chng minh l mang li nhiu ch li v sc khe cho con ngi. Cc hp cht Anthocyanin c hp thu vo trong d dy dng phn t (Passamonti, Vrhovsek v Mattivi, 2002) hoc c th c h tr bi mt c ch vn chuyn qua mt. Ngoi ra, phn t Anthocyanin cng khng b bin i di tc dng ca h vi khun trong rut non. V th, phn t Anthocyanin cng khng thay i trong huyt tng v nc tiu. Cc nghin cu gn y cho thy cc Anthocyanin ch c hp thu mc rt thp, ch khong 0,016% n 0,11% lng tiu th ngi.

Mc d ch c c th hp thu mt lng rt nh, cc phn t Anthocyanin sau khi c chuyn ha c th biu hin nhng hot tnh nh chng ung th, chng x va ng mch, chng vim, gim mc thm thu, v ca mao mch, c ch s ng t ca cc tiu huyt cu v thc y s to thnh cytokine t iu ha cc phn ng min dch. Tt c nhng hot tnh ny u da trn kh nng chng oxy ha ca cc Anthocyanin. Cng nh kh nng ny, cc hp cht Anthocyanin cn gip bo v mng d dy chng li nhng thng tn do s oxy ha, v vy hon li giai on u ca bnh ung th d dy, ung th rut v rut k. Hot tnh v thnh phn ca h vi khun ng rut c th b thay i sau khi dng cc dch trch t cc loi qu mng (berry) c cha cc hp cht flavonoid gm c Anthocyanin. Mt s hot tnh sinh hc ca cyanidin-3-glucoside: Gim kh nng b oxy ha ca LDL Bo v hng cu khi cc tc ng ca s oxy ha. Gim s phn tch DNA. c ch c tnh ca cc i thc bo bng cch gim hm lng NO.

Ngn chn tc hi ca tia UV trong liposome. Khng vim. Chng cc bnh tim mch. c ch s tc ng v pht trin ca cc t bo ung th. 9.2.1. Hot tnh chng oxy ha: S oxy ha v sc khe con ngi: S to thnh qu mc cc gc t do c th lm vt qu kh nng chng oxy ha ca cc enzyme nh glutathione peroxidase, catalase, superoxide dismutase, v cc hp cht chng oxy ha nh glutathione, tocopherol hay acid ascorbic. Hu qu l cc protein, lipid v DNA s l mc tiu tn cng ca cc gc t do, lm tn thng n cc enzyme, mng t bo v cc vt cht di truyn. Cc gc t do v cc nhm oxy hot ng c lin quan n mt s ri lon thn kinh trong c th. Ngoi ra, s oxy ha cc lipoprotein t trng thp l nguyn nhn chnh thc y cho cc chng bnh tim mch vnh v bnh x va ng mch. Tuy nhin,t bo cng c nhng h enzyme v cc cht chng oxy ha gip chng li nhng tc ng xu bi cc gc t do. Cc cht chng oxy ha t nhin c trong thc phm

(nh tocopherol v acid ascorbic) cng c th vai tr quan trng gip ngn chn s hnh thnh cc gc t do. Mt s c ch chng oxy ha ca Anthocyanin: S thiu electron t nhin ca cc phn t Anthocyanin gip cho cc hp cht ny c bit hot ng. Mt s c ch chng oxy ha ca Anthocyanin c c t cc nghin cu nh: Ngn chn cc gc hot ng bng cch cho hydro. Chelate cc ion kim loi xc tc cho cc phn ng oxy ha Lin kt vi cc protein, to phc cht bn. Phn ng th vo 2 nhm hydroxy nm v tr ortho trong vng B ca Anthocyanin v cyanidin ng vai tr quan trng gip n nh cc gc t do. Ngoi ra, nhm ortho- dihydroxy ny cn c kh nng chelate cc ion kim loi v t ngn chn s peroxy ha lipid. Tsuda v cng s tm cch gii thch c ch chng oxy ha ca Cyanidin-3- glucoside (C3G) bng cch cho phn ng vi gc alkylperoxyl. Da trn sn phm ca phn ng, cc tc gi cho rng C3G chng oxy ha theo c ch khc vi alpha-tocopherol v gi thuyt rng C3G ph v cu trc v qut cc gc t do. Cc hp cht flavanoid ni chung trong c Anthocyanin chng oxy ha bng cch qut cc gc O2 t do, hay phn ng vi cc gc peroxy tham gia vo phn ng oxy ha dy chuyn. Nh vo kh nng cho cc gc t do H+, cc hp cht ny c th c ch c phn ng peroxy ha lipid. Cc yu t nh hng n kh nng chng oxy ha ca Anthocyanin: Rt nhiu cc nghin cu cho thy mi tng quan kh cao gia hm lng mu trong nguyn liu (tri cy, rau) vi kh nng chng oxy ha. Ngoi ra, s khc nhau v cu trc gia cc Anthocyanin cng c th nh hng n hot tnh chng oxy ha. Trn thc t, bng cch thay i v tr v loi gc ha hc gn vo vng thm ca phn t Anthocyanin, kh nng nhn electron c thn t cc gc t do cng s khc nhau.

Hnh 12: S to phc gia cyanidin v DNA

Theo kt qu nghin cu trn cc Anthocyanin v aglycon cyanidin ca chng trong cherry cht, kh nng c ch s oxy ha lipid ca cyanidin cao hn cc glucoside ca n, t cho thy rng hot tnh chng oxy ha ca Anthocyanin l do aglycon quyt nh. S lng cc gc ng v tr C3 cng c vai tr rt quan trng n kh nng chng oxy ha, s lng cc phn t ng cng t kh nng chng oxy cng cao hn. Bng phng php o kh nng hp th gc oxy thy rng cyanidin 3 glucoside c hot tnh ORAC cao nht trong s 14 loi Anthocyanin c kim tra v cao hn 3,5 ln hot tnh ca Trolox (cht tng ng vitamin E). Chnh v vy, ngi ta ang hng n kh nng s dng nhiu phng php khc nhau trong qu trnh nui cy huyn ph t bo hoc chn ging thc vt nhm thu c loi Anthocyanin nh mong mun.

9.2.2.Hot tnh chng ung th: Tt c cc cn bnh ung th u do s hnh thnh, tng trng v suy vong ca cc t bo bt bnh thng. Cc khi u l do s tch t ca cc t bo vi s lng ln hn nhu cu cn thit cho s pht trin, sa cha v hot ng ca cc m. Trong cc nghin cu in vitro v in vivo, cc hp cht Anthocyanin u cho thy kh nng gim s tng trng ca cc t bo ung th v c ch s hnh thnh khi u mt cch ng k. C ch chng ung th ca Anthocyanin ni ring v cc hp cht phenolic ni chung vn cha c xc nh chc chn, c th lin quan n kh nng c ch cc enzyme cyclooxygenase v hot tnh chng oxy ha. Mt s nghin cu v kh nng chng ung th ca Anthocyanin nh: Cc Anthocyanin trong khoai lang tm v bp ci tm c ch s ung th rut kt trong chut Cc aglycon c trong nhng loi Anthocyanin ph bin nht nh cyanidin, delphinidin, maldivin, pelargonidin v petunidin u c kh nng c ch s pht trin ca cc t bo ung th d dy, rut kt, phi, lng ngc ngi. Theo nghin cu ny, nhm hydroxy t do v tr C3 trong dng cu trc flavylium ca Anthocyanidin gp phn c ch s pht trin ca cc dng t bo ung th ca ngi c kho st. Trong Anthocyanin, nhm hydroxy v tr C3 lun b th bi cc gc ng, v v vy, cc Anthocyanin khng c c kh nng c ch nh trn. Ngoi ra, s nhm hydroxyl v methoxyl trong vng B ca cyanidin cng nh hng rt ln n kh nng c ch cc dng t bo ung th c nghin cu. Hot tnh c ch cao nht thuc v maldivin, vn c nhm hydroxy v tr 3 v 4 v nhm methoxy cc v tr 3 v 5. Nghin cu v kh nng c ch s di cn ca cc t bo ung th ca Anthocyanin. S di cn i hi t bo phi c kh nng di ng, kh nng kt dnh b mt v s hot ng ca cc protease ngoi bo nh serine protease, matalloproteinase (MMPs), cathepsine, nhm phn hy mng li ngoi bo (extracellular matrix ECM), cho php cc t bo di cn. Trong , cc enzyme quan trng nht l serine protease v MMPs. MMPs l nhm enzyme phn hy ph thuc vo Zn c kh nng phn hy cc protein ca mng li ngoi bo nh collagen, proteoglycan, fibronectin v elastin. C MMP 2 v MMP 9 u xut hin nhiu trong nhiu khi u c tnh v gp phn vo s di cn. Cc hp cht cyanidin 3-rutinoside and cyanidin 3-glucoside chit sut

t qu du tm c chng minh l c kh nng c ch cc enzyme MMP, hn ch s di cn ca dng t bo ung th c nghin cu.

9.2.3. Hot tnh chng cc bnh tim mch: Cc hp cht flavanoid ni chung v cc Anthocyanin ni ring c cho l c kh nng lm gim nguy c mc bnh ng mch vnh bi kh nng ngn chn s oxy ha cc lipoprotein c t trng thp (LDL) trong huyt tng. S oxy ha cc hp cht ny c xem nh mt bc quan trng trong s hnh thnh cc khi x ng mch v t dn n cn bnh ng mch vnh. Vai tr ca Anthocyanin trong vic phng chng cc bnh tim mch c lin quan trc tip n hot tnh chng oxy ha, gim vim, tng bn v kh nng thm thu ca thnh mch mu, c ch s ng t ca cc tiu huyt cu. Kt lun: lun Cho n nay, phn ln cc nghin cu v vai tr ca Anthocyanin i vi sc khe con ngi vn cha th gii thch r rng v cc c ch hot ng sinh hc, s hp th, s phn b trong c th v trong cc m ca Anthocyanin. Mt trong nhng vn phc tp nht lc ny l trong c th cht mu Anthocyanin khng khi no hot ng ring l. Bt c loi thc vt no cha Anthocyanin u c cha c rt nhiu nhng hp cht flavonoid khc. C n hn 4000 hp cht flavonoid khc nhau dng v loi nhm th. Chnh v vy, khi vo bn trong c th, cc hp cht ny c th phi hp vi nhau lm tng hot tnh sinh hc hay mc hp thu. V th, nu sn xut cc chit xut thc vt ch cha Anthocyanin, hot tnh sinh hc thu c s khng th so snh vi cc dch trch t thc vt, vi rt nhiu hp cht phi hp vi nhau. Mt vn khc l s khng n nh ca cc cht mu Anthocyanin. V vy, khi p dng mt s phng php truyn thng trch ly v phn tch cc thnh phn khi dch trch th, cc phn t Anthocyanin c th b phn hy v b v hot trong cc bc tinh sch tip theo. V vy, cc nghin cu nhm xc nh hoat tnh sinh hc ca cc dch trch t thc vt rt kh nh gi ngun gc ch thc ca hot tnh sinh hc c loi thc vt .

Trong thin nhin, rt nhiu cc hp cht flavonoid, trong c c Anthocyanin c tng hp l do cc tc nhn kch thch vt l v ha hc t mi trng, thng thng l mt yu t gy stress. V vy, khi mun gia tng s tng hp Anthocyanin mt loi thc vt, cn phi to c mt kch thch tng t nh cc tc nhn kch thch t mi trng. iu ny rt kh thc hin trong t nhin nhng c th c thc hin d dng hn trong mi trng nui cy c bit (nh cc huyn ph t bo). II. Cc nguyn liu c cha Athocyanin:1. Bp ci tm.

1.1. Tng quan: Bp ci c tn khoa hc l Brassica oleracea. var. capitata. Lizg. Bp ci tm (Brassica oleracea var capitata ruba) bt ngun t ty bc Chu u. n khong gia th k 16 bp ci tr thnh loi rau quan trng Chu u. T chu u bp ci tm c a i trng rng ri nhiu ni trn th gii. Ti Vit Nam bp ci tm c trng nhiu Lt. Tuy nhin, vic sn xut bp ci tm khu vc nhit i v nhit i cn mc hn ch v bp ci tm thch hp vi nhng vng ni cao hoc ni c ma ng lnh. o Bp ci tm sinh trng tt nht khu vc c nhit trung bnh ngy 15-20 C, nhit o dao ng nhit ngy v m l 5 C. Vng nhit i ch gp iu kin ny ni c cao o trn 800m so vi mc nc bin. Nhit vt qu 25 C bp ci vn pht trin nhng kh nng cun bp hn ch. V th Vit Nam bp ci tm c trng nhiu Lt. Bng phng php ti u ho thc nghim trong qu trnh chit tch, xc nh c hm lng Anthocyanin trong bp ci tm Lt chim khong 1,11%. Sc t chnh c chit xut t bp ci tm l cyanidin 3,5-diglucoside ca h mu anthocyanin v c mu sc thay i r rt theo pH mi trng. Trong mi trng acid n c mu bn v khi mi trng chuyn sang base mu ca n chuyn sang xanh v n nh trong thi gian di. y l c tnh rt khc so vi cc anthocyanin ca mt s nguyn liu khc (mu thay i lin tc ti pH kim) nh l ta t, qu du,...

Hnh 13: Bp ci tm

Hnh 14: Cng thc cu to ca Cyan idin 3,5 diglucoside

1.2.Thnh phn ha hc: Bng 2: Thnh phn ha hc ca bp ci Thnh phn Vitamin K Vitamin C Dietary fiber Mangan Vitamin B6 (pyridoxine) Folate Acid bo omega- 3 Vitamin B1 (thiamin) Vitamin B2 (riboflavin) Caci Kali Vitamin A Tryptophan Protein Magie Hm lng 73.35 mcg 30.15 mg 3.45 g 0.18 mg 0.17 mg 30.00 mcg 0.17 g 0.09 mg 0.08 mg 46.50 mg 145.50 mg 198.00 IU 0.01 g 1.53 g 12.00 mg

(Ngun: www.whfoods.com)

Bp ci l thc phm u tin c pht hin l gy ra b n h bu c khi c dng trong thnh phn thc phm cho th n hng ngy trong mt cuc nghin cu v bnh giang mai vo nm 1928. Khm ph ny sau c cc nh nghin cu khc xc nhn vo cc nm 1929, 1933. Bp ci thu hoch vo ma ng c tc dng mnh hn loi thu hoch vo ma H, v tc dng ny gia tng khi bp ci c hp chn..v sau cc nh nghin cu ghi nhn nu cho th n thm iodine trong khu phn hng ngy chung vi bp ci, d sng hay hp chn, th th s khng b bu c. nguyn liu ny, hm lng Athocyanin c xc nh bng phng php trch ly. Gii thiu v phng php trch ly: Trch ly l qu trnh tch mt hoc mt s cht tan trong cht lng hay trong cht rn bng mt cht lng khc gi l dung mi. Nu qu trnh tch cht ha tan trong cht lng bng mt cht lng khc th gi l trch ly lng lng. Nu qu trnh tch cht ha tan trong cht rn bng mt cht lng th gi l trch ly rn lng. trng hp ny ta s dng trch ly lng - lng. Qu trnh trch ly cht lng gm 2 giai on:Giai on u: l giai on trn ln, phn tch 2 pha vo vi nhau to s tip

xc pha tt cho dung cht truyn t hn hp u vo dung mi. Nu thi gian tip xc pha th qu trnh truyn vt cht xy ra cho n khi t cn bng gia hai pha.Giai on k tip: l giai on tch pha, hai pha tch ra d dng hay khng

ty thuc vo s sai bit khi lng ring gia hai pha. Mt pha l pha trch gm ch yu dung mi v dung cht, mt pha gi l pha rafinat gm ch yu phn cn li ca hn hp ban u. thng th cc cu t trong hn hp v dung mi u t nhiu ha tan vo nhau v th trong hai pha u c s hin din ca c ba cu t. Cc phng php trch ly theo on: Trch ly mt on: qu trnh c th thc hin lin tc hay gin on.

Hn hp ban u v dung mi c trn ln vi nhau to thnh hn hp, qu trnh truyn khi din ra gia hai pha c tin hnh cho n khi h cn bng sau hn hp lng v tch pha.

Hnh 15: Qu trnh trch ly mt on. Trch ly nhiu on giao dng: y l s ko di ca qu trnh trch ly mt on trong pha rafinat i lin tc qua mi on c tip xc vi dung mi mi. Qu trnh cng c th thc hin lin tc hay gian on.

Hnh 16: Qu trnh trch ba on giao dng

Trch ly lin tc nhiu on nghch dng: Nguyn liu v dung mi i ngc chiu nhau. Nguyn liu i vo u ny cn dung mi i vo u kia. Hai pha trch v pha rafinat lin tc i ngc chiu nhau qua mi on cui cng c hai dng sn phm. Qu trnh ny cn t on trch hn qu trnh nhiu on giao dng vi cng 1 lng dung mi hoc cn t dung mi hn vi cng s on trch.

Hnh 17: Qu trnh trch nhiu on nghch dng

Trch ly lin tc nghch dng c hon lu: trong qu trnh trch ly lin tc nghch dng, dng sn phm trch c nng ln nht khi t cn bng vi dng nhp liu. Vic s dng dng hon lu ca pha trch lm pha trch c nng cao hn. Cc yu t nh hng n qu trnh trch ly: Thc cht qu trnh trch ly l qu trnh khuch tn, v vy s chnh lch nng gia 2 pha chnh l ng lc ca qu trnh. Khi chnh lch nng ln, lng cht trch ly tng, thi gian trch ly gim ta thc hin bng cch tng t l dung mi so vi nguyn liu. Din tch tip xc gia nguyn liu vi dung mi: cn tng din tch tip xc gia nguyn liu vi dung mi bng cch nghin nh. N cn lm v cu trc t bo, thc y qu trnh tip xc trit gia dung mi v vt liu. Tuy nhin, kch thc v hnh dng ca vt liu sau khi lm nh cng c gii hn v nu chng qu mn s b lng ng ln lp nguyn liu, tc cc ng mao dn kh trch ly. Tnh cht ca vt liu cng nh hng ln n hiu sut trch ly nh m ca nguyn liu.

Nhit c tc dng tng tc khuch tn v gim nht, phn t cht ha tan chuyn ng d dng khi khuch tn gia cc phn t dung mi, tuy nhin nhit l yu t gii hn, v khi nhit cao qu c th xy ra cc phn ng khc khng cn thit gy kh khn cho cc qu trnh cng ngh... Thi gian trch ly cng nh hng n hiu sut trch ly. Khi thi gian tng ln lng cht khuch tn tng nhng thi gian phi c gii hn, khi t c mc trch ly cao nht nu ko di thi gian s khng mang li hiu qu kinh t. 1.3.Quy trnh cng ngh chit tch Anthocyanin t bp ci tm:

Hnh 18: S quy trnh cng ngh chit tch Anthocyanin t bp ci tm.

Tin hnh th nghim: Nguyn liu Ha cht s dng: Bp ci tm HCl 1% CH3COONa Dng c - thit b: My o quang ph pH k T lnh ng Thit b sy chn khng Phng php th nghim: Th nghim 1: Nghin cu nh hng ca nhit bc m v m nguyn liu n kh nng chit tch v bn mu anthocyanin Mc ch th nghim: Tm ra nhit bc m v m thch hp cho hiu sut trch ly cao nht v bn mu nht. Chun b th nghim: Nguyn liu bp ci tm c ra sch Dung mi Ethanol nc - HCl (vi t l 1:1 v 1%HCl) B tr th nghim: Th nghim c b tr hon ton ngu nhin vi 2 nhn t v 3 ln lp li. Nhn t A l nhit bc m, bao gm: 0 o A1 l nhit 10-15 C ( bc hi nc trong t lnh) 0 o A2 l nhit 40 C Ethanol KCl

0 o A3 l nhit 50 C Nhn t B l m sau khi bc m, bao gm: o B1 l m 15% ( 0,5%) o B2 l m 20% ( 0,5%) o B3 l m 25% ( 0,5%) Mu i chng nguyn liu khng sy C S nghim thc 3x3+1= 10 mu

A1

A2

A3

B1

A1B1

A 2 B1

A3B1

B2

A1B2

A 2 B2

A2B3

B3

A1B3

A 2 B3

A3B3

o A1B1: Mu bc m 10-15 C m 15% o A1B2: Mu bc m 10-15 C m 20% o A1B3: Mu bc m 10-15 C m 25%

o A2B1: Mu bc m 40 C m 15% o A2B2: Mu bc m 40 C m 20% o A2B3: Mu bc m 40 C m 25% o A3B1: Mu bc m 50 C m 15% o A3B2: Mu bc m 50 C m 20% o A3B3: Mu bc m 50 C m 25%

Phng php nghin cu: Phng php ha hc - chit tch v lm sch anthocyanin Phng php pH vi sai * Xc nh hm lng anthocyanin th Ti pH = 1 cc anthocyanin tn ti dng oxonium hoc flavylium c hp th cc i Ti pH = 4,5 chng dng carbinol khng mu. o hp th cc i ca 2 mu pH = 1 v pH = 4,5 ti bc sng ca hp th cc i 515 nm, so vi hp th ti bc sng 700 nm ( c ca mu) % anthocyanin ton phn =A.M.k.V .100% .l.m(100 W).10 2

(2.1)

Trong : A: mt quang ( hp th ca anthocyanin) A = (A515(pH=1) - A700(pH=1)) - (A515(pH=4,5) - A700(pH=4,5))

vi

A515(pH=1): hp th cc i ti bc sng 515 nm A700(pH=4,5): hp th cc i ti bc sng 700 nm M: khi lng phn t ca anthocyanin khng k ion Cl- hay nc tinh th ha : hp th phn t. l: chiu dy cuvet, cm V: th tch anthocyanin thu c, l k: pha long m: khi lng nguyn liu ban u, g W: m nguyn liu, % * Xc nh mu: xc nh mu ca anthocyanin ta tin hnh o mt quang A pH = 1 ti bc sng max

v 420 nm (vng mu tm ca nh sng hp th) mu = (2.2)

A max A 420

Kt qu nghin cu: nh hng ca h dung mi n hm lng anthocyanin Sau khi chit tch v lm sch anthocyanin trong cc h dung mi trn, din trn hnh 19. nh hng ca h dung mi n mu anthocyanin: tin hnh o mt

quang A, p dng cng thc (2.1) thu c kt qu hm lng anthocyanin bng 3 v biu

Tin hnh th nghim tng t, nhng y o hp th cc i ca anthocyanin bc sng 515 nm v 420 nm v p dng cng thc (2.2), ta tnh c mu ca anthocyanin c c t cc dung mi khc nhau. Kt qu mu v mu sc cm nhn c bng mt cng c th hin bng 3 v hnh 19.

Bng 3: nh hng ca dung mi n hm lng v mu anthocyanin Dung mi Ch s Hm lng, % mu Mu cm nhn Methanol HCl 0,949 3,267 en xanh Ethanol - Nc - hydrochloric acid 0,492 3,032 vng xanh HCl 1,046 3,134 en xanh -

- nc - nc

Nc formic acid

- Nc acetic acid

-

0,452 2,825 vng xanh

0,107 2,059 vng nht xanh

bng mt Mi trng kim

Hnh 19: th biu din nh hng ca cc h dung min hm lng v mu anthocyanin trong qu trnh

T kt qu thu nhn c hnh 19, bng 3 cho thy thy rng h dung mi ethanol - nc HCl cho hm lng anthocyanin ln v mu tng i cao. Hn na ethanol t c hn so vi methanol v l dung mi thng dng, r tin, d kim. y cng l mt trong nhng l do m ta chn h dung mi ethanol - nc - HCl trong quy trnh cng ngh chit tch v lm sch anthocyanin th. H dung mi ny cng c th dng tch cht mu anthocyanin t cc loi rau qu tng t. nh hng ca t l dung mi (ethanol-nc) n hm lng anthocyanin:

Tin hnh kho st nh hng ca t l dung mi (ethanol - nc) n hm lng anthocyanin trong du. Cc thng s ban u v kt qu c biu din bng 3, th hnh 20. T cho thy t l dung mi ethanol - nc l 1:1 (vi 1% HCl) cho hiu sut chit cao nht.

Bng 4: T l dung mi v hm lng anthocyanin % Stt Vethanol 30 2 3 4 5 6 7 8 9 35 40 45 50 55 60 65 70 Vnc 70 65 60 55 50 45 40 35 30 3: 7 7: 13 2: 3 9: 11 1: 1 11: 9 3: 2 13: 7 7: 3 0,869 0,895 0,974 1,019 1,047 1,025 0,957 0,893 0,828 % T l % anthocyanin

Hnh 20: nh hng ca t l dung mi (ethanol -nc) n hm lng anthocyanin Kt lun Chn c h dung mi tt cho qu trnh chit tch cht mu anthocyanin c mu cao l ethanol - nc - HCl.

Tm c t l dung mi ethanol-nc l 1:1 (vi 1% HCl) hm lng anthocyanin thu c nhiu nht.

1. Go np than:1.1. Tng quan:

Go np than l mt loi go c bit c trng nhiu Nam B. V th m ru np than c sn xut ch yu Vng Nam B. Go np than gm 4 loi: Np cm c Ha Np en Khnh Vnh Np than Long t la lc Np cm Cc loi la ny c nng sut khng cao, thng ch t 2.8 - 3.3 tn/ha. Hin nay dn vng ng Bng Nam B phn loi np than theo mu sc ht cch ny, np than c chia thnh 2 loi: Np than en huyn Np than hng Mu ca Np than l mu ca Anthocyanin. go. Theo

Hnh 21: Ht Np than

1.2. Thnh phn ha hc:

Bng 5: Thnh phn ha hc ca go np than Thnh phn Hm lng

Nc Protein Lipid Glucid Acid hu c

14 8.2 1.5 74.9 0.6

Tro

0.8

Nhm nghin cu t i hc bang Louisiana (M) phn tch mu cm ly t go np cm. H pht hin thy chng cha hm lng rt cao cht chng oxi ha Anthocyanin mt cht c tim nng chng li bnh ung th, tim mch v nhiu bnh khc.

2.3.Qu trnh trch v tinh ch Athocyanin t go np than.

Go (50g) Dung mi ngm (18:85) 100ml Ngm trong cc dung dch (15:85) (t nht 3 ngy nhit phng) Dung mi ngm (18:85) 100ml Lc ra Trn cc phn trch Cho qua dng kh CO2 Ly dch trch th Qua ct nha trao i ion Acid 5% Ra vi acid 5%(50ml)

(50ml)

Ra vi C2H5OH Cho qua dng CO2 Dch tinh khit

1.3. Nhn xt v kt lun:

Nh ni trn, hp thu ca Athocyanin ph thuc vo dung mi trch, c th nh:

i vi dung mi trch l cc loi acid (HCl, CH 3COOH, HCOOH ) v nc th hp thu t gi tr cc i ti = 510nm i vi dung mi l cc loi acid (HCl, CH3COOH, HCOOH ) V C2H5OH th hp thu t gi tr cc i ti 535nm =

Qua ta thy, cng mt thi gian ti ph nht nh th trong dung mi m c C2H5OH th hp thu t gi tr cao hn so vi trong dung mi gia acid vi nc, do hm lng Athocyanin trch c nhiu hn, nguyn nhn l do mu ca Athocyanin tan c trong cn. Mt khc, trong cc loi acid th dung mi gia acid CH3COOH v C2H5OH l c hp thu cao nht (A=1.4) trong khi dung mi trch l HCl v C2H5OH th ch c ( A = 0.855 ). V vy, trong qu trnh trch chit Athocyanin th dung mi CH 3COOH v C2H5OH l dung mi trch tt nht. 1. Du ty:

3.1. Tng quan:

Du ty c tn khoa hc l Fragaria hay cn gi l du t l mt chi thc vt ht kn thuc h hoa hng cho qu c nhiu ngi a chung. Du ty c con ngi bit n t nm 1400 chu u do ngi La M pht hin v c trng vo thi Trung c.

Hnh 22: qu du ty Du ty cha nhiu cht chng oxy ha, gm cc Flavonoid: Anthocyanin, catechin, quercetin, kaempferol..v cc vitamin nh A, B1, B2, C, trong hm lng vitamin C kh cao, y l nhng tnh u vit ca qu du ty gip tng sc khng, chng nhim trng, nhim c, cm cm v chng stress. 3.2. Thnh phn ha hc: Bng 6: Thnh phn dinh dng ca qu du ty. Thnh phn Hm lng

Nng lng Nc protein lipid glucid vitamin A

46 kcal 84 g 1.8 g 0.4 g 7.7 g 5 g

Beta caroten Vitamin E Vitamin B1 Vitamin B2 Vitamin pp Vitamin B6 Vitamin C Calcium( Ca ) kalium( k) natrium( Na ) Tro

30 g 0.58 g 0.03 g 0.06 g 0.3 g 0.06 g 60 g 22 mg 190 mg 0.7 mg 0.8 g

3.3. Phng php nghin cu: phng php ny, Anthocyanin th c chit t qu du trong nc SO2. Da vo s khc nhau ca cc nng SO2, nhit v chn ca nguyn liu ta c th thu c Anthocyanin vi cc hm lng khc nhau cng vi mt s c tnh: mu, bn mu, hm lng mu.ca chng. 3.3.1. Nguyn liu: Qa du tm. 3.3.2. Phng php: Xc nh hm lng Anthocyanin theo phng php PH vi sai. 3.3.3. Tin hnh th nghim: Da theo chn ca qu du:

Qa du trc tin c loi b tp cht v nhng qu dp nt, sau phn loi da theo chn v tin hnh o mu bng my o Minolta, xc nh hm lng Anthocyanin bng phng php vi sai, kt qu thu c nh sau: Bng 7: Kt qu hm lng Anthocyanin theo chn ca qu du. Loi du M t trng thi Hm lng Anthocyanin (%) Cha chn Gn chn Va chn Qu chn Mu hng nht, cng Mu hng, cng Mu tm en,cng Mu en sm, mm 0.147 0.474 0.994 1.287

Vi bng kt qu trn, ta thy nhng qu du qu chn th hm lng Anthocyanin thu c l cao nht. Da theo nng SO2: Tin hnh chit Anthocyanin th vi cc dung mi c nng SO 2 l 500ppm, 700ppm, 900ppm, 1100ppm, 1300ppm v 1500ppm, c b sung 1% HCl nhit 30oC, thi gian chit l 30 pht. Thu c kt qu hnh sau:

Hnh 23: Kt qu hm lng Anthocyanin cc nng SO2 khc nhau. Vi kt qu ny ta thy, hm lng Anthocyanin thu c cao nht nng SO2 l 1100ppm. Da theo nhit chit: Tin hnh kho st ti cc nhit chit khc nhau: 30oC, 40oC, 50oC, 60oC, v 70oC, nng SO2 l 1100ppm, thi gian chit l 30 pht. Sau khi phn tch v tnh ton hm lng Anthocyanin, ta thu c kt qu theo hnh sau:

Hnh 24: Kt qu hm lng Anthocyanin cc nhit khc nhau. 3.3.4. Kt qu nghin cu v cc c tnh ca Anthocyanin: Mt trong nhng c tnh quan trng ca Anthocyanin l bn mu ca Anthocyanin. Theo kt qu nghin cu cho thy bn ny thay i ph thuc vo loi dung mi, ph, nhit v thi gian chit. nghin cu bn mu ca Anthocyanin tch t qu du ngi ta tin hnh o hp thu ti bc sng hp thu cc i ca mu trc v sau khi gia nhit. bn mu c th hin bng phn trm mu cn li so vi ban u, xc nh bi cng thc:

% Mu cn li = AA1.100% Trong : A: l hp thu ca mu ti thi im o xc nh A1: l hp thu ca mu ti bc sng hp thu cc i ban u vi mi mu nghin cu. Sau qu trnh nghin cu v tnh ton ta thu c cc kt qu trong bng sau:

Thi gian Nc SO2 Ban u 15 pht 30 pht 45 pht 60 pht 100

Ph = 3 Etanol nc 100 96.36 95.37 89.79 85.50

Ph = 4 Nc SO2 100 96.36 94.47 91.63 88.66 Etanol nc 100 94.47 91.50 86.81 82.57

Ph = 5 Nc SO2 100 95.84 92.91 90.71 86.30 Etanol nc 100 92.65 91.76 88.82 85.59

Ph = 6 Nc SO2 100 94.82 90.46 88.56 83.65 Etanol nc 100 91.30 87.62 84.95 80.26

97.54 96.21 94.12 91.61

Bng 8: Phn trm mu cn li theo thi gian 95oC ti cc PH khc nhau ca Anthocyanin th chit trong dung mi nc sulfite v etanol/nc.

Nhn xt: Khi gia nhit n 95oC cc dung mi v PH khc nhau th bn mu ca Anthocyanin gim dn theo thi gian v theo chiu tng ca PH. cng nhit v PH bn mu ca Anthocyanin chit trong nc SO2 cao hn khi chit trong etanol/nc. 95oC Anthocyanin tng i bn nhit khi gia nhit trong khong thi gian 60 pht, sau 60 pht Ph = 3 phn trm mu ca Anthocyanin chit trong nc SO2 l 91.61%, trong khi chit trong etanol/nc l 85.50%.Ti ph = 6 phn trm mu cn li l thp nht, sau 60 pht gia nhit l 80.26%. So vi cc gi tr Ph cn li th PH = 3 Anthocyanin t b thoi bin do nhit hn. 3.4. Kt lun: Qua kt qu th nghim cho thy nhng qu du qu chn th hm lng Anthocyanin cao hn so vi cc qu du cha chn. V vy, thu c hm lng Anthocyanin cao th ta nn thu hi nhng qu du va chn hoc qu chn, cn nhng qu cha chn hoc gn chn th khng nn thu hi v hm lng Anthocyanin trong chng cn thp.

So vi cc nng khc th ti nng SO2 1100ppm v nhit 60oC cho hm lng Anthocyanin tng i cao. Nguyn nhn l do nc c nng SO2 thp, t bo ca nguyn liu dng u trng, thm ch c th gy ra s hp th nc, khi tip tc tng nng n mt gi tr nht nh ty thuc vo c im ca nguyn liu th lc t bo s trng n cc i v v ra ng thi gii phng cc cht mu nm trong phn khng bo. Tuy nhin nu tip tc tng nng SO2 th trng thi nhc trng li hnh thnh lm cho t bo b mt nc ng thi cng li, lm gim kh nng thm ca mng t bo dn n Anthocyanin vn b gi nguyn trong phn khng bo v qu trnh chuyn khi xy ra kh khn hn. V nhit , khi nhit tng th nht gim ng thi tng vn tc khch tn do to iu kin thun li cho vic chuyn ng ca cht ha tan t nguyn liu vo dung mi v vy m hm lng Anthocyanin tng dn. Trong cng ngh thc phm nhit l yu t gii hn v khi nhit tng, cc thnh phn c trong thc phm d b bin i. Do Anthocyanin c tnh cht c trng l tan tt trong nc m nn hm lng Anthocyanin t cao nht ti 60oC l d nhin. Cn 70oC tr ln th nhit cao dn nn cc Anthocyanin b bin tnh, do hm lng Anthocyanin bt u gim dn. hn ch hin tng bin tnh do nhit n mc thp nht ngi ta thng chit nhit thng. V dung mi ta thy hm lng Anthocyanin v mt mu ca n trong dung mi etanol/nc cao hn so vi chit trong nc SO 2. Nguyn nhn l do trong dung dch, phn t Anthocyanin dng monomer v polymer, nc SO2 trong qu trnh tip xc vi nguyn liu s kh mt phn Anthocyanin dng monome do lm cho mt mu gim i ng thi cng lm gim hm lng Anthocyanin. III. Cc cht Anthocyanin thng mi:1. Encolor lipid DS ( cng ty Reggianna, Antociani, Italy ) l dch trch v nho c c. 2.Redberry liqid 88-01 c sn xut bi cng ty Gilette Food Inc, l dch trch qu ca

alderberry, blackberry, blueberry, blackcurrant. 3.Sambucus NR 54 c sn xut bi cng ty Gilette Food Inc,l dch trch anthocyanin t elderberry. 4.San Red Liqid RC-EX c sn xut bi cng ty San Ei Chemical Industries ( Nht ) l dch c dc t bp ci m chng cha anthocyanin diacyl ha. 5.Powdered San Red PC c sn xut bi cng ty San Ei Chemical Industries l bt sy kh ca dch tr ch t bp ci .

6.Powdered San Red PC c sn xut bi cng ty San Ei Chemical Industries l bt sy kh

ca dch trch t bp ci tm.III. Phng php nghin cu, bo v mu ca Anthocyanin:

1. Phng php nghin cu: S nghin cu: Ngmc GiiNguyn C st ph hp 1%HPLC Bin